Table 1.

Kinetic parameters for BcPMH wild type at pH 7.5

Substrate		kcat (s-1)	KM (mM)	kcat/KM (s-1 M-1)	kcat/kuncat†,‡	(kcat/KM)/kuncat†,§ (M-1)	(kcat/KM)/kw†,¶	KTS∥ (M)
Phosphate monoester	1a*	(2.1 ± 0.2) ∗ 10-4	0.33 ± 0.03	0.63 ± 0.08	106.2	109.7	1011.5	1.9 ∗ 10-10
	1b	(7.7 ± 0.1) ∗ 10-3	0.35 ± 0.02	22 ± 1	105.2	108.7	1010.4	2.2 ∗ 10-9
Phosphate diester	2a	2.12 ± 0.02	0.071 ± 0.004	(3.0 ± 0.2) ∗ 104	1013.7	1017.9	1019.6	1.4 ∗ 10-18
	2b	5.8 ± 0.1	0.63 ± 0.04	(9.2 ± 0.6) ∗ 103	1013.4	1016.6	1018.3	2.8 ∗ 10-17
Phosphate triester	3b	> 3.7 ∗ 10-5	> 2.4	(1.6 ± 0.1) ∗ 10-2	> 102.9	105.5	107.2	3.2 ∗ 10-6
Phosphonate monoester	4a	1.58 ± 0.04	1.23 ± 0.09	(1.3 ± 0.1) ∗ 103	1012.9	1015.8	1017.6	1.4 ∗ 10-16
	4b	2.73 ± 0.06	0.19 ± 0.02	(1.5 ± 0.1) ∗ 104	1011.2	1014.9	1016.7	1.2 ∗ 10-15
Sulfate monoester	5a*	(1.0 ± 0.1) ∗ 10-4	58 ± 5	(1.7 ± 0.3) ∗ 10-3	108.4	109.6	1011.3	2.5 ∗ 10-10
	5b	(4.0 ± 0.1) ∗ 10-2	68 ± 4	0.59 ± 0.04	107.6	108.7	1010.5	1.9 ∗ 10-9
Sulfonate monoester	6a*	(7.0 ± 0.3) ∗ 10-4	0.51 ± 0.09	1.4 ± 0.2	106.7	1010.0	1011.7	1.0 ∗ 10-10
	6b	(1.2 ± 0.1) ∗ 10-2	0.24 ± 0.03	49 ± 7	106.3	109.9	1011.7	1.1 ∗ 10-10

^*K_M was determined as the competitive inhibition constant (K_ic) for the conversion of phosphate diester 2b. By using this K_ic as the K_M value for that substrate, the k_cat could be derived (see SI Text).

^†Values for k_uncat and k_w are listed in Table S1.

^‡First-order rate enhancement.

^§Catalytic proficiency.

^¶Second-order rate enhancement.

^∥K_TS, the transition state binding constant, was calculated as the inverse of the catalytic proficiency [(k_cat/K_M)/k_uncat].